Automatic multiple-decanting centrifuge and method of treating physiological fluids

ABSTRACT

A centrifuge is capable of holding a sample container in selected orientations, either during or after centrifugation, to drain supernatants between two or more chambers of the container. The draining may be gravity or centrifugal draining. This allows an automated process to subject a sample to a first physical or chemical treatment to produce a first supernatant, the first supernatant to be subjected to a second physical or chemical treatment, and a second supernatant to be separated from a desired component.

CROSS REFERENCE TO RELATED APPLICATION

This application is a division of U.S. application Ser. No. 08/435,662,which was filed on May 5, 1995, now U.S. Pat. No. 5,707,331.

TECHNICAL FIELD

This invention relates to the art of automatic centrifugation. Inparticular, the invention relates to apparatus and procedures usingautomatic, multiple decanting with centrifugation. In a preferredembodiment, an automated procedure separates blood components andproteins including the separation of fibrinogen from blood.

BACKGROUND

The separation of components through centrifugation is well known. Forexample, in the medical field it is common to subject a sample of bloodto centrifugation to produce a precipitate of cellular material and asupernatant of plasma. The plasma is then decanted to complete theseparation of these components.

U.S. Pat. Nos. 5,178,602 (Wells) and 5,047,004 (Wells) show an automatedcentrifuge, which includes structure for holding a centrifuge tube,after centrifugation, in a position that allows the supernatant to drainfrom the tube and into another container by gravity. The holdingstructure shown in these patents comprises a locking mechanism mountedfor axial movement with respect to the axis of rotation of thecentrifuge. An electromagnet that is easily controlled causes the axialmovement.

It is also known to decant a supernatant by the process of centrifugaldraining. According to that process, a centrifuge rotates a centrifugetube while the tube is held in a position such that the supernatant isdrained from the tube by centrifugal forces.

Fibrin sealants for treating wounds are known and are typically producedby combining a fibrinogen/Factor XIII component with bovine thrombin.When these are mixed, a fibrin tissue adhesive results, which is appliedto the wound. Descriptions of compositions for use as tissue sealantsare given in U.S. Pat. Nos. 5,292,362 and 5,209,776 (Bass et al.). Thefibrinogen is obtained from plasma, either pooled or autologous, andcryoprecipitation is one known technique for separating fibrinogen fromplasma. One cryoprecipitation technique is described in U.S. Pat. Nos.5,318,524 and includes the centrifugation of thawing plasma to produce aprecipitate containing fibrinogen/Factor XIII. Other techniques forproducing fibrinogen/Factor XIII include inducing precipitation of thecomponent by addition of such agents as Ammonium Sulfate or polyethyleneglycol (PEG) to blood plasma.

SUMMARY OF THE INVENTION

Several known chemical procedures include repeated steps of physicalseparation between two or more components. Separation based on densitydifferences between the components is often by centrifugation, and theresulting supernatant is decanted to complete the separation. Each stepprovides an opportunity for error, which would be reduced by automationof the process.

In accordance with the invention, chemical procedures requiring severalcentrifugation steps are automated, to reduce the time required by aclinician and eliminate the potential for errors. Apparatus inaccordance with the invention includes a multiple-chamber container anda centrifuge designed to receive the container and subject its contentsto predetermined centrifugation steps as well as gravity and centrifugaldecanting of the supernatant.

A preferred container in accordance with the invention includes firstand second chambers separated by an intermediate wall. The first chamberis designed to receive a first liquid, such as human blood. The secondchamber is located adjacent the first chamber, and the wall between thechambers is such that a supernatant in the first chamber will flow overthe top of the wall and be drained into the second chamber by gravitywhen the container is held in the proper orientation. The supernatant inthe second chamber may then be subjected to a mixing action and then maybe subjected to a second centrifugation. The container can also be heldin a second position whereby a second supernatant is caused to flow backover the wall into the first chamber by centrifugal forces resultingfrom a second centrifugation.

A centrifuge in accordance with the invention includes a rotatablesupport with a swinging frame for receiving the multiple-chambercontainer and means for locking the container in either of at least twopositions for draining supernatant fluids from the chambers. Preferably,the locking means is an electro-magnetically operated disk mounted formovement axially with respect to the axis of rotation of the rotatablesupport. The centrifuge is preferably operated under the control of anelectronic circuit, which may include a programmed array logic (PAL) orother circuitry, that causes the rotor to operate in accordance with apredetermined program and controls the locking means such that it locksthe container in predetermined orientations in conjunction withoperation of the rotor.

While many different programs for operation of the centrifuge can bedeveloped, depending on the desired results, a preferred operation isfor the production of autologous fibrinogen. Prior techniques forproduction of fibrinogen require several distinct steps, each of whichrequires a skilled technician but does not eliminate an opportunity forerror. These steps include separation of plasma from cellularcomponents, treatment of the plasma with a precipitating agent, andseparation of a fibrinogen precipitate “pellet” from the plasma. Theseparation of plasma from blood and the separation of the fibrinogenpellet from plasma typically require centrifugation first of the bloodand then of the plasma, with addition of at least one precipitatingagent between the steps. Thus, the production of fibrinogen in the priorart has been complex and error-prone.

In accordance with this embodiment of the invention, a volume of thepatient's anticoagulated blood is placed in the first chamber of thedisposable container, and a precipitation agent is placed in the secondof the chambers. The container is then placed in the swinging frame ofthe centrifuge, and the control circuit is activated to initiate theoperation of the centrifuge. The centrifuge first rotates the containerfor a time period that has been determined to be adequate for separatingthe cellular components from the supernatant plasma. During this time,the swinging frame will have rotated outwardly substantially due tocentrifugal forces on the container. While the frame is in the outwardlyrotated position, the locking means is activated to lock it there. Therotation of the support is then terminated. As the rotational velocityof the support decreases, the supernatant fluid, being no longer subjectto the centrifugal forces, flows out of the first chamber and into thesecond chamber by gravity. The cellular component is more viscous and,thus, flows toward the second chamber at a rate less than that of theplasma. Preferably, however, a divider in the form of a disk is placedin the first chamber to restrict the flow of the cellular components andplasma below the disk. The disk is at a depth that provides apredetermined volume of plasma, which is normally near the expectedboundary between the supernatant and cellular components. After a periodof time that has been determined to allow an adequate amount of theplasma to flow into the second chamber, the locking means is deactivatedto release the container, whereby it assumes an upright position withthe cellular component remaining in the first chamber and the plasma nowin the second chamber. The rotatable support is then alternatelyactivated and deactivated for short intervals to mix the plasma with theprecipitating agent in the second chamber. Interaction between theprecipitating agent and the plasma initiates precipitation of fibrinogenand Factor XIII from the plasma. The support is then again rotated toaccelerate the precipitation of the fibrinogen/Factor XIII and to createa pellet in the bottom of the second chamber. As a final step, thelocking means is again activated to lock the container in a positionsuch that the supernatant resulting from precipitation of the fibrinogenis decanted by centrifugal draining into the first chamber. In thisstep, the container is held substantially upright, and the support isrotated to apply centrifugal forces to the supernatant, whereby it flowsover the wall between the chambers and into the first chamber. Thelocking means is then inactivated, the container removed from thecentrifuge, and the fibrinogen/Factor XIII removed from the secondchamber for further processing. In a preferred embodiment, thefibrinogen/Factor XIII is reconstituted, and then combined withthrombin, and applied to a patient to treat a wound.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a perspective of a container and centrifuge in accordance withthe invention.

FIG. 2 is a vertical cross section of a preferred embodiment of acontainer.

FIGS. 3a and 3b are partial vertical cross sections of the centrifuge ofFIG. 1.

FIGS. 4a through 4f are schematic diagrams illustrating a preferredmethod of operation of the centrifuge of the invention.

DETAILED DESCRIPTION OF THE INVENTION

With reference to FIGS. 1 and 2 of the drawings, a centrifuge 2 isdesigned to receive a container 4 in accordance with the invention. Thecentrifuge is capable of subjecting the container to a series of stepsthat will be described in detail below. The container includes at leasttwo chambers, 6 and 8. Chamber 6 is designed to receive a first fluid tobe treated, such as blood. Chamber 8 is designed to receive fluids thathave been decanted from chamber 6, such as a supernatant plasmaresulting from centrifugation of blood in chamber 6.

A preferred form of the container is shown in detail in FIG. 2. Asshown, the container comprises three primary parts. A base part ispreferably molded and includes the chambers 6 and 8 and a bridge 7,which connects the two chambers. A lid 11, also preferably molded, fitsover the tops of the chambers to close them. The lid includes cup shapedextensions 12 and 14, each of which is centrally aligned with arespective one of the chambers 6 and 8. Extension 12 has a access portin the form of centrally located opening 13, while extension 14 has acentrally located opening 15. The openings receive syringe needles topermit fluids to be injected into the chambers or withdrawn therefrom.Membranes 16 and 17 cover the openings 13 and 15 to maintain sterility.The membranes are preferably heat sealed into the extensions 12 and 14during construction by providing a cavity for receiving the membranes.After a membrane is inserted, the upper edges of the cavity are foldedover and welded, e.g., ultrasonically, to retain the membrane.

The lid also includes a bridge 7′ that cooperates with bridge 7 in thebase to form a fluid channel 18, connecting chambers 6 and 8. As shown,the bridge 7 extends above the tops of the chambers 6 and 8 to preventcommunication between the chambers by “splashing.” Intentional fluidcommunication between the two chambers will be described in detailbelow.

A separation disk 20 is preferably placed in chamber 6 near, but alwaysabove, the expected vertical position of the boundary betweensupernatant plasma and cellular components after a first centrifugationof a blood sample. The hematocrit is known to vary among individuals,and the exact amount of plasma that will result from a blood samplecannot be accurately predicted without prior testing of the sample.Thus, disk 20 is located such that the plasma above the disk aftercentrifugation of a predetermined volume of blood is a predeterminedvolume of plasma. The upper surface of the disk 20 is tapered toward anedge, and the edge includes at least one groove 22 that allows fluidcommunication between the parts of the chamber 6 that are above andbelow the disk 20.

In a preferred embodiment, a cylindrical support 24 is attached to thelower surface of the disk to set the location of the disk duringassembly.

A hollow tube 26 is provided to facilitate introduction of the bloodsample to the portion of the chamber 6 that is below the disk 20. Thetube 26 extends from just below the opening 13 through disk 20. Thus, asyringe needle inserted through opening 13 pierces membrane 16 andcommunicates with tube 26 to allow injection of the blood sample intothe bottom of the chamber 6. The groove 22 permits downward movement ofthe plasma and cellular components during centrifugation but retardsmovement of the cellular components during decanting. Also, an air vent27 is provided for chamber 8 to facilitate introduction and withdrawalof fluids.

In use, a container 4 is placed in a holder on the rotor of thecentrifuge as indicated in FIG. 1. To balance the rotor, two suchcontainers are preferably placed in the centrifuge in diametricallyopposed positions. Of course, only one container may be used and aweight or “dummy” container used to balance the rotor.

FIGS. 3a and 3b are partial cross sections of a preferred embodiment ofa centrifuge showing the container locked in two different positions. Arotor shaft 28 is connected to a motor (not shown), which rotates theshaft. A rotor 30 is mounted to the shaft for rotation and has a frame32 pivotally mounted to the rotor 30 at pivot connection 34. The topsurface (not shown) of the frame 32 has two circular openings forreceiving the chambers 6 and 8 whereby the container can be placed inthe frame such that the contents of the container will be subjected tocentrifugal forces as the rotor is rotated. A bias spring 35 ensuresthat the frame 32 will pivot to an upright position when centrifugationis terminated. The frame 32 may also be shaped to reduce windresistance, as known in the art.

A locking plate 36 is mounted coaxially with the shaft 28 for engagingthe frame 32 to lock the container in desired orientations. The plateand the mechanism for controlling the positions of the plate may be thesubstantially the same as that shown in my previous U.S. Pat. No.5,178,602. For example, an electromagnet 38 may be provided to controlthe position of the locking plate by action on a permanent magnet 40,which is attached to the locking plate.

Preferably, the electromagnet 38 and magnet 40 are positioned such thatthe locking plate can be placed in either of two positions. In a firstposition, shown in phantom lines, the plate does not engage the frame32, and the frame 32 is free to rotate about pivot 34. In a secondposition, shown in solid lines at 36′, the locking plate engages one oftwo parts of the frame 32 to hold it in one of two selectedorientations. In the position shown in FIG. 3a, a lip of the plateengages a protuberance 42 on the frame 32 to lock the container in theorientation shown in FIG. 3a. In the position shown in FIG. 3b the plate36 engages an upper edge of the frame 32 to lock the container in thetilted position shown in FIG. 3b. The locking plate preferably rotateswith the rotor whereby it can be moved to engage the frame duringcentrifugation of the contents of the container.

The operation of the centrifuge in a preferred embodiment of theinvention will be described with regard to FIGS. 4a through 4f. In afirst step, blood is introduced into chamber 6 of the container throughopening 13. The blood has preferably been obtained from a patient, butit may be pooled or obtained from another. A precipitating agent 43,e.g., PEG, is then placed in chamber 8, preferably by injection throughopening 15. The container with blood and precipitating agent are thenplaced in the centrifuge for automated operation.

In the first step of automated operation, the container is allowed toswing freely as the blood is subjected to centrifugation. As illustratedin FIG. 4a, the cellular component 44 of the blood will be separatedfrom the plasma component 46 in this step. After a predetermined timeperiod, e.g., five minutes, the locking plate 36 is moved to a positionshown at 36′ whereby the container 4 is held in the position shown inFIGS. 3b and 4b, and rotation of the rotor is stopped. In this position,the plasma component 46 flows through channel 18 by the force ofgravity. The chamber is held in the position of FIG. 4b for preferablyabout 3 seconds, which is adequate to allow the plasma to drain bygravity into the chamber 8 but is not so long that the more viscouscellular component 44 drains into the chamber 8. The plasma 46 andprecipitating agent 43, which was previously placed in chamber 8, arenow both in chamber 8. To provide complete mixing of these fluids, thelocking plate is lowered, and the rotor is caused to accelerate anddecelerate alternately for 10-20 seconds, as illustrated in FIG. 4c. Theprecipitating agent causes the fibrinogen/Factor XIII to separate fromthe plasma, and this separation is assisted by centrifuging the contentsof the container a second time. This second centrifugation may be for aperiod of about five minutes. A fibrinogen pellet 48 is, thus, formed inthe bottom of the chamber 8, as illustrated in FIG. 4d. At this stage ofthe process, the plasma supernatant 46 remains in chamber 8.

Plasma 46 is separated from the fibrinogen pellet 48 by stoppingrotation of the centrifuge rotor to allow the container to pivot to theupright position shown in FIGS. 3a and 4e. The locking plate 36 is thenactivated to lock the container in that orientation by engagement withprotuberance 42, and the container is again rotated by the rotor for aperiod of about three to eight seconds. This rotation causes thesupernatant plasma 46 to flow back through channel 18 and into chamber 6by centrifugal draining, as illustrated in FIG. 4e. Thus, the fibrinogenpellet and plasma have now been separated. As a final step, thecontainer is subjected to another centrifugation illustrated in FIG. 4ffor about fifteen seconds, whereby the fibrinogen pellet is forced intothe bottom of the chamber 8.

The automated process for production of fibrinogen is at this pointcomplete, and the fibrinogen pellet is preferably extracted from thecontainer 8 by a syringe for further processing. For example, thefibrinogen may be reconstituted and combined with thrombin to produce asealant or an adhesive.

The apparatus of the invention may be used for other automatedprocesses. For example, another technique for the separation offibrinogen from blood in accordance with the structure of the inventionuses cryoprecipitation. According to this technique, plasma is frozen toa temperature of about minus 20° C., thawed, and then centrifuged toseparate the fibrinogen from plasma. The multiple-decanting apparatus ofthis invention may be used to automate cryoprecipitation by inclusion ofa temperature control device 50 in thermal contact with the centrifuge.The temperature control device may comprise any of several knownstructures, including liquid nitrogen or liquid oxygen based devices andrefrigeration devices.

To effect automated cryoprecipitation, a sample of blood is placed inthe first chamber 8, and the container is then placed in the centrifugeand subjected to a first centrifugation. The plasma is then drained intothe second chamber 8, for example by gravity draining. The temperaturecontrol device is then activated first to freeze the plasma and then toallow the plasma to thaw. The thawed plasma is subjected to a secondcentrifugation, which separates fibrinogen from the remainder of theplasma. The supernatant plasma is then separated from the fibrinogen bydraining it back into the first chamber, for example by centrifugaldraining, whereby only fibrinogen remains in the second chamber. Thecontainer is then removed from the centrifuge, and the fibrinogenremoved from it for use as described above. Of course, thefreeze-thaw-centrifuge process may be carried out any number of timesbefore the supernatant is drained back into the first chamber.

Modifications within the scope of the appended claims will be apparentto those of skill in the art.

1. A method for automatic separation of components from fluidscomprising placing first and second chambers in a centrifuge, subjectingsaid first chamber to centrifugation, locking said chambers in firstpositions such that a supernatant in said first chamber drains into saidsecond chamber, subjecting said second chamber to centrifugation, andlocking said chambers in second positions for allowing a supernatant insaid second chamber to transfer to another of said chambers.
 2. A methodaccording to claim 1 wherein said another of said chambers is said firstchamber, said supernatant in said first chamber drains into said secondchamber by gravity draining, and said supernatant in said second chambertransfers into said first chamber by centrifugal transfer.
 3. A methodaccording to claim 1 further comprising the step of freezing saidsupernatant in said second chamber prior to said step of subjecting saidsecond chamber to centrifugation.
 4. A method according to claim 3further comprising thawing said supernatant and wherein said step ofsubjecting said second chamber to centrifugation is performed as saidsupernatant is thawing.
 5. A method according to claim 4 wherein saidanother of said chambers is said first chamber, said supernatant in saidfirst chamber drains into said second chamber by gravity draining, andsaid supernatant in said second chamber transfers into said firstchamber by centrifugal transfer.
 6. A method for separation ofcomponents of a substance comprising: placing a first substance in afirst chamber of a container having at least two separate chambers influid communication with each other, rotating said container tocentrifuge said first substance and separate said first substance into afirst component and a second component, locking said container in afirst position that allows said first component to flow into a secondchamber of said container, rotating said container again to centrifugesaid first component to produce a third component and a fourthcomponent, and locking said container in a second position that allowssaid third component to flow to said first chamber.
 7. A methodaccording to claim 6 wherein said first component drains into saidsecond chamber by gravity.
 8. A method according to claim 7 furthercomprising the step of centrifugally transferring said third componentby rotating said container while locking said container in said secondposition.
 9. A method according to claim 8 wherein said first substancecontains blood, said first component contains plasma, and said fourthcomponent contains fibrinogen.
 10. A method according to claim 9 whereinsaid second chamber is supplied with a precipitating agent prior to saidstep of rotating said container to centrifuge said first substance. 11.A method according to claim 10 wherein said precipitating agent is PEG.12. A method for centrifuging substances comprising: providing aremovable container having a plurality of chambers for receivingsubstances to be centrifuged; placing one or more substances in saidcontainer; rotating said container a first time to subject saidsubstances to centrifugation; locking said container in a first positionto allow a supernatant in one of said chambers to transfer into a secondof said chambers; and locking said container in a second position androtating said container a second time to transfer a supernatant in saidsecond chamber to said one of said chambers.
 13. The method of claim 12,wherein the step of locking said container in said first position causessaid supernatant in said one of said chambers to transfer substantiallyinto said second chamber by gravity.
 14. The method of claim 12, whereinthe step of locking said container in said second position and rotatingsaid container causes a supernatant in said second chamber to transfersubstantially into said one of said chambers by centrifugaltransferring.
 15. The method of claim 12, wherein the step of lockingthe container in said first position comprises holding said container insaid first position for a predetermined period of time.
 16. The methodof claim 12, wherein the step of locking the container in said firstposition comprises controlling the position of a movable plate.
 17. Themethod of claim 12, further comprising controlling the locking androtating of said container to provide automatic multiple decanting,wherein the container is locked and/or rotated at respective intervalsof predetermined duration.
 18. The method of claim 12, furthercomprising the step of mixing said one or more substances in saidcontainer by accelerating and decelerating the rotation of thecontainer.
 19. The method of claim 12, further comprising the step ofmaintaining the substances in at least one chamber separate from eachother with a divider.
 20. The method of claim 19 wherein said dividerhas an opening for allowing said substances to be discharged from saidat least one chamber.
 21. The method of claim 12, wherein the step ofplacing one or more substances into said container comprises the step ofplacing blood in said one of said chambers and a precipitating agent insaid second of said chambers, wherein the step of rotating saidcontainer a first time causes a supernatant plasma to be separated froma cellular component of said blood, and the step of locking saidcontainer in said first position causes said supernatant plasma to besubstantially transferred from said one of said chambers into saidsecond of said chambers, while substantially leaving said cellularcomponent in said one of said chambers.
 22. The method of claim 21,further comprising the step of mixing said supernatant plasma and saidprecipitating agent in said second chamber, and rotating said containeragain to cause fibrinogen and Factor XIII to be precipitated from thesupernatant plasma to create a pellet in said second of said chambers.23. The method of claim 22, wherein the step of locking and rotatingsaid container a second time causes a supernatant resulting from saidprecipitation to be substantially transferred from said second chamberto said one of said chambers, thereby leaving behind said pellet in saidsecond chamber.
 24. A method for centrifuging substances comprising:providing a unitary container having a plurality of chambers therein forreceiving substances to be centrifuged; placing one or more substancesinto said container; rotating said container a first time to subjectsaid substances to centrifugation; locking said container in a firstposition to allow a supernatant to be transferred from one chamber toanother chamber by gravity; locking said container in a second positionand rotating said container a second time to cause a supernatant to betransferred from one chamber to another chamber by centrifugal transfer.25. The method of claim 24, wherein the container comprises a first anda second chamber, wherein the step of placing substances within thecontainer comprises placing one substance in the first chamber and asecond substance in the second chamber.
 26. The method of claim 25,wherein the step of rotating said container a first time causes asupernatant to separate from the one substance in said first chamber,wherein the step of locking the container in said first position causesthe supernatant in said first chamber to be transferred by gravity intosaid second chamber through a passage between said first and secondchambers.
 27. The method of claim 26, further comprising the step ofmixing said supernatant and second substance in said second chamber byaccelerating and decelerating the rotation of the container for apredetermined time, wherein said mixing helps to produce a precipitationin said second chamber.
 28. The method of claim 27, further comprisingrotating the container again to accelerate the formation of saidprecipitation in said second chamber, wherein the precipitate in saidsecond chamber is forced to the bottom of said second chamber in theform of a pellet.
 29. The method of claim 28, wherein the step ofrotating the container a second time causes the supernatant resultingfrom said precipitation to be transferred from said second chamber tosaid first chamber, leaving behind the precipitation in the form of apellet in said second chamber.
 30. The method of claim 29, furthercomprising controlling the steps in the process to provide automaticmultiple decanting which allows for activation of one or more steps inthe process for a predetermined period of time.
 31. The method of claim30, wherein the step of placing one or more substances in said containercomprises placing blood in said first chamber and a precipitating agentin said second chamber.
 32. A method for treating physiologicalproducts, comprising: providing a centrifuge; providing a containerhaving at least a first chamber and an adjacent second chamber, whereineach of the first and second chambers has a top portion, a bottomportion and a set of walls, wherein the top portions of the firstchamber and second chamber are adjacent each other and connected by abridge for transferring fluid therebetween when said container is in apredetermined orientation; providing a holder assembly attached to thecentrifuge and effective to removably receive the container, wherein theholder assembly is effective to orient the container in saidpredetermined orientation; and placing a physiological product in one ofsaid chambers.
 33. The method of claim 32, wherein the chambers includelid portions, thereby forming a closed container.
 34. The method ofclaim 33 wherein at least one of the chambers includes an access portfor transference of a liquid.
 35. In a method of treating physiologicalfluids, the improvement comprising providing a container adapted tocontain said fluids during treatment, wherein said container comprises:a first sterile chamber having a first top portion, a first bottomportion and a first set of walls; a second sterile chamber adjacent saidfirst sterile chamber and having a second top portion adjacent saidfirst top portion, a second bottom portion and a second set of walls; abridge connecting said first top portion of the first chamber and saidsecond top portion of the second chamber, such that a fluid can betransferred from the first chamber to the second chamber while thecontainer is positioned at a predetermined angle, and means for steriletransfer of a fluid to or from at least one of said chambersindependently of the other of said chambers and located near the top ofat least one of said chambers, and placing a physiological fluid in oneof said chambers.
 36. The method of claim 35, wherein the chambersinclude a lid portion.
 37. A method for treating physiological productsand maintaining sterility of said products during said treatingcomprising: providing a container having a plurality of closed, sterilefluid-receiving chambers, a bridge forming a fluid path allowing fluidcommunication between a first of said chambers and a second of saidchambers when said container is in a predetermined orientation, and atleast one access port allowing sterile access to at least one of saidchambers, providing a centrifuge having a holder removably receivingsaid container and allowing said container to assume a first orientationwherein a physiological product in one of said chambers is subjected tocentrifugation and said predetermined orientation wherein fluid in saidfirst of said chambers flows along said fluid path to said second ofsaid chambers, and said centrifuge comprises a locking element thatselectively holds said container in said predetermined orientation, andplacing a physiological product in one of said chambers.
 38. A methodaccording to claim 37 wherein said holder comprises a frame pivotallymounted to a rotor of said centrifuge.
 39. A method according to claim37 wherein said locking element comprises a movable locking plate thatis movable between free and locking positions, wherein said movablelocking plate allows said container to assume said first orientationwhen in said free position and holds said container in saidpredetermined position when in said locking position.
 40. A methodaccording to claim 39 wherein said centrifuge further comprises anelectromagnet for moving said locking plate to one of said locking endfree positions.
 41. A method according to claim 37 wherein said holdercomprises a frame pivotally mounted to a rotor of said centrifuge, andsaid locking element comprises a movable locking plate that is movablebetween free and locking positions, wherein said movable locking plateallows said container to assume said first orientation when in said freeposition and holds said container in said predetermined position when insaid locking position.
 42. In a method of treating physiological fluids,the improvement comprising providing a container adapted to contain saidfluids during treatment, wherein said container comprises a base forminga plurality of sterile chambers, each of said chambers having a bottomand a top, a bridge connecting top portions of at least two of saidchambers and arranged to provide a sterile fluid channel from a first ofsaid at least two sterile chambers to a second of said at least twosterile chambers when said container is in a predetermined orientation,a lid closing said top of each of said plurality of chambers, and anaccess port near the top of at least one of said chambers that allowssterile transfer of a liquid to or from said at least one of saidchambers independently of the other of said chambers from the exteriorof said container to the interior of said at least one of said chambers,and placing a physiological fluid in one of said plurality of sterilechambers.
 43. A method according to claim 42 wherein said plurality ofsterile chambers and said bridge comprise a molded base part.
 44. Amethod according to claim 43 wherein said container is substantiallyrigid.
 45. A method according to claim 42 wherein said container furthercomprises a separation disk in one of said chambers.
 46. A methodaccording to claim 42 wherein said plurality of chambers comprise firstand second adjacent chambers having adjacent sidewalls and said bridgeis formed at the tops of said adjacent sidewalls.